JP5212969B2 - Chroman compound and process for producing the same - Google Patents

Description

  The present invention relates to a chroman compound having a strong whitening action and an antioxidant action and having high stability, and a method for producing the same.

  The cause of skin blackening is melanin pigment accumulated in the skin. As a whitening agent that inhibits the synthesis of melanin pigments, various compounds have been developed as active ingredients in cosmetics, foods, and pharmaceuticals. Arbutin, kojic acid, lucinol, ascorbic acid and derivatives thereof have been developed as compounds showing a whitening action. Whitening agents such as arbutin, kojic acid, and lucinol inhibit the activity of tyrosinase, a key enzyme in melanin synthesis, thereby inhibiting melanin synthesis and preventing skin blackening. Ascorbic acid and its derivatives inhibit melanin accumulation by reducing melanin and intermediates in melanin production.

  However, these whitening agents all have insufficient whitening action, and development of more effective whitening agents is expected.

  An antioxidant is a substance excellent in a reducing action that inhibits oxidation of a substance. This reduction action can prevent oxidation of other substances. Antioxidants are capable of eradicating radicals such as active oxygen and lipid peroxides such as superoxide radicals and hydroxy radicals that cause severe damage to living bodies and cause inflammation, aging, and carcinogenesis. Moreover, in the biosynthetic pathway in melanin synthesis, the antioxidant exhibits a reducing action on melanin and intermediates in melanin production, and also inhibits melanin synthesis.

  Several compounds have been developed as antioxidants so far. Representative compounds that have been developed so far include butylhydroxytoluene (BHT) and α-tocopherol (vitamin E). In particular, α-tocopherol exhibits excellent antioxidant activity and is an antioxidant that is frequently used as an active ingredient in cosmetics, foods, and pharmaceuticals. However, α-tocopherol and butylhydroxytoluene are oil-soluble compounds that are insoluble in water, and are difficult to formulate into aqueous preparations. Existing antioxidants also lack stability and cause coloration, discoloration, and odor changes. In addition, many existing antioxidants are insufficiently effective. Therefore, development of more effective and stable water-soluble antioxidants is expected.

  The present inventors have clarified that mycelia of Daedalea dickinsii produce a relatively large amount of a compound having the following molecular structure (daedalin A), and have a water-soluble property that has both a whitening effect and an antioxidant effect. daedalin A is disclosed as a new type of whitening agent, antioxidant and the like (Patent Document 1).

  Since this daedalin A is derived from Horotakutake, it can be safely applied to the human body. However, this daedalin A is stable when compared with conventional active ingredients such as whitening agents, but it has been clarified that it turns yellow upon long-term storage. Since higher stability is desired for compounds used in the components of external skin products such as cosmetics, it is more stable and more effective than this daedalinA, and is effective in whitening, antioxidant and active oxygen removal. There is a need for materials having

  In addition, as a method for producing daedalinA, the present inventors have disclosed a method for separating it from the metabolites of mycelia of Horotakutake.

  As another production method, Non-Patent Document 2 discloses a method for producing a racemate of daealin A by organic synthesis. However, the production method disclosed in Non-Patent Document 2 is not practical because the synthesis yield is low and the production procedure is complicated. Therefore, a more efficient synthesis method is required.

Prior art document information relating to the invention of this application includes the following.
JP2006-124386 SynlettNo. 2. 322-324 (2002)

  An object of the present invention is to provide a compound that is more stable than daedalinA and has a whitening action, an antioxidant action, an active oxygen removal action, and the like. Another object of the present invention is to provide a method for efficiently organically synthesizing daedalinA or a compound of the present invention.

  The present inventors have started development for improvement of the production method of daedalin A by the mycelium of B. edulis, improvement of the functionality of daedalin A by chemical conversion, and establishment of a practical method for synthesizing daedalin A racemate.

  As a result, it was found that daedalin A is a substance that is produced during the growth process of mycelia of Horotake mushroom and is metabolized (converted) to other substances according to the growth. Therefore, in order to efficiently produce daedalinA using Borotake, it is necessary to separate the mycelium of Borotake from the product of metabolites after culturing the mycelium of Borotake for a certain period of time before the structure of daedalin A is converted to other substances. is there.

  In addition, it was found that a compound having the following molecular structure hydrogenated to the double bond at the 3rd and 4th positions of daedalinA can provide a whitening agent and an antioxidant superior in stability and functionality to daedalin A.

さらには、daedalinA は有機合成によって得られる daedalin A のラセミ体においても daedalin A と同等の抗酸化作用、美白作用を示すことを見出すとともに、非特許文献１において開示されている既存の合成方法よりも優れたdaedalin A の合成方法を新たに開発し、本発明の完成に至った。

Furthermore, daedalin A has been found to exhibit the same antioxidant and whitening effects as daedalin A in the racemic form of daedalin A obtained by organic synthesis, and moreover than the existing synthesis method disclosed in Non-Patent Document 1. A new method for synthesizing daedalin A has been newly developed, and the present invention has been completed.

  Accordingly, the present invention provides the following.

  (Item 1) The following structural formula:

を有する化合物。

A compound having

  (Item 2) A whitening agent containing the compound according to item 1.

  (Item 3) An antioxidant containing the compound according to item 1.

  (Item 4) An active oxygen scavenger containing the compound according to item 1.

  (Item 5) A skin external preparation containing the compound according to Item 1.

  (Item 6) The following structural formula:

に表される化合物のラセミ混合物、または、
以下の構造式：

A racemic mixture of the compounds represented by
The following structural formula:

に表される化合物のラセミ混合物を含有する、美白剤。

A whitening agent comprising a racemic mixture of the compound represented by

  (Item 7) The following structural formula:

に表される化合物のラセミ混合物、または、
以下の構造式：

A racemic mixture of the compounds represented by
The following structural formula:

に表される化合物のラセミ混合物を含有する、抗酸化剤。

An antioxidant comprising a racemic mixture of the compounds represented by

  (Item 8) The following structural formula:

に表される化合物のラセミ混合物、または、
以下の構造式：

A racemic mixture of the compounds represented by
The following structural formula:

に表される化合物のラセミ混合物を含有する、活性酸素除去剤。

An active oxygen scavenger containing a racemic mixture of the compounds represented by

  (Item 9) The following structural formula:

に表される化合物のラセミ混合物、または、
以下の構造式：

A racemic mixture of the compounds represented by
The following structural formula:

に表される化合物のラセミ混合物を含有する、皮膚外用剤。

A skin external preparation containing a racemic mixture of the compound represented by

  (Item 10) The following structural formula:

に表される化合物を製造する方法であって、以下：
（ａ）ホウロクタケ (daedalea dickinsii) の菌糸体を10日から50日間培養する工程、
を包含する、方法。

A method for producing a compound represented by the following:
(A) a process of culturing mycelia of daedalea dickinsii for 10 to 50 days;
Including the method.

  (Item 11) The following structural formula:

に表される化合物を製造する方法であって、以下：
ａ）ホウロクタケ(daedalea dickinsii) の菌糸体を10日から50日間培養する工程；
ｂ）工程（ａ）の培養物から、以下の構造式

A method for producing a compound represented by the following:
a) culturing the mycelium of daedalea dickinsii for 10 to 50 days;
b) From the culture of step (a), the following structural formula

に表される化合物を抽出する工程；および、
ｃ）工程（ｂ）で抽出された化合物に水素を付加する工程、
を包含する、方法。

Extracting a compound represented by: and
c) adding hydrogen to the compound extracted in step (b);
Including the method.

  (Item 12) The following structural formula:

に表される化合物のラセミ混合物を製造する方法であって、以下：
（ａ）パラトルエンスルホン酸と以下の構造式

A process for preparing a racemic mixture of the compounds represented by
(A) Paratoluenesulfonic acid and the following structural formula

に表される化合物とを、1,2-ジクロロエタン中で反応させる工程、
を包含する、方法。

A step of reacting the compound represented by: in 1,2-dichloroethane,
Including the method.

  (Item 13) The following structural formula:

に表される化合物のラセミ混合物を製造する方法であって、以下：
（ａ）パラトルエンスルホン酸と以下の構造式

A process for preparing a racemic mixture of the compounds represented by
(A) Paratoluenesulfonic acid and the following structural formula

に表される化合物とを、1,2-ジクロロエタン中で反応させる工程、
（ｂ）工程（ａ）で生成された化合物に水素を付加する工程、
を包含する、方法。

A step of reacting the compound represented by: in 1,2-dichloroethane,
(B) adding hydrogen to the compound produced in step (a);
Including the method.

  The compound of the present invention has tyrosinase inhibitory activity, whitening action, antioxidant action, active oxygen removal action and the like, and is very stable. Therefore, tyrosinase activity inhibitors, whitening agents, antioxidants, active oxygen (free radical) scavengers and the like can be prepared using the compounds of the present invention. Furthermore, since the compound of the present invention also has an anti-inflammatory action, it is useful as an anti-inflammatory agent. The action of inhibiting tyrosinase activity inhibits melanin synthesis, resulting in a whitening effect. The free radical scavenging action prevents skin aging and is effective in improving skin wrinkles and tarmi.

  According to the present invention, an excellent compound as an active ingredient such as a whitening agent, an antioxidant, an active oxygen scavenger, an anti-inflammatory agent, and a tyrosinase inhibitor is provided. Therefore, the present invention provides an excellent whitening agent, antioxidant, active oxygen scavenger, anti-inflammatory agent, and tyrosinase inhibitor. In addition, an efficient method for producing this active ingredient is provided.

  The present invention will be described below. Throughout this specification, it should be understood that the singular forms also include the plural concept unless specifically stated otherwise. In addition, it is to be understood that the terms used in the present specification are used in the meaning normally used in the art unless otherwise specified. Thus, unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification, including definitions, will control.

(Definition of terms)
As used herein, the term “fruit body” is a reproductive body that produces a spore in a fungus and may be used interchangeably with the term “spore body”. Ascomycota and basidiomycetes refer to various forms of mycelial tissue, such as ascomycetes and basidiomycetes.

  As used herein, the term “mycelium” is used interchangeably with the term “mycelium” and is the basic structure that constitutes the nutrients of filamentous fungi, and includes multicellular and multinuclear ones. Mycelium develops from the germ tube of the spore and extends by tip growth.

  As used in the present invention, the term “effective amount” refers to a compound of the present invention such as a whitening effect, an antioxidative effect, an active oxygen scavenging effect, and the like, when ingested or administered to a human. The amount that produces a significant effect. Typically, an effective amount of a compound of the invention is 0.1 mg to 100 mg, preferably 0.2 mg to 50 mg, more preferably 0.5 mg to 30 mg, and even more preferably 0.8 mg per administration. -20 mg, most preferably 1 mg-10 mg, but these may vary depending on the intended effect and the condition of the subject.

(Experimental method)
In the present invention, the whitening action was confirmed by a tyrosinase inhibition test and a melanin production inhibition test using B16 melanoma cells.

  The tyrosinase inhibition test is a method for measuring the tyrosinase inhibitory activity of a test substance from the change in absorbance. Tyrosinase catalyzes the oxidation reaction between tyrosine and DOPA (3,4-dihydroxyphenylalanine), the rate-limiting reaction of melanin synthesis that causes pigmentation. Therefore, this substance that inhibits tyrosinase activity inhibits melanin synthesis, resulting in a whitening effect.

  The melanin production inhibition test using B16 melanoma cells is a test method for measuring the melanin production inhibition effect by B16 melanoma cells of the test substance. Active melanin synthesis is carried out in melanoma cells, and substances that inhibit this melanin synthesis show an excellent whitening effect.

  The antioxidant action of the present invention was evaluated by a DPPH radical scavenging rate measurement test. 1,1-diphenyl-2-picrylhydrazyl (hereinafter abbreviated as DPPH) used in the DPPH radical scavenging rate measurement test has a stable free radical in the molecule. In the DPPH radical elimination rate measurement, the free radical elimination rate is measured by measuring the absorbance. Substances having DPPH radical scavenging activity can be expected to remove singlet oxygen, hydroxy radicals, and lipid peroxide radicals, as well as prevent inflammation, aging such as skin wrinkles and tarmi, and canceration of cells.

  Further according to the invention, the following structural formula:

に表される化合物、ならびに、以下の構造式：

As well as the following structural formula:

および

and

に表される化合物のラセミ混合物、あるいは、これらを１種または２種以上を、皮膚外用剤および食品、医薬品に配合することで優れた美白剤、抗酸化剤、活性酸素除去剤を提供することができる。また、本発明に従って、これらの化合物を効率的に生産することも可能となる。

A racemic mixture of the compounds represented by the above, or a combination of one or more of these in skin preparations for external use, foods and pharmaceuticals to provide an excellent whitening agent, antioxidant and active oxygen remover. Can do. In addition, according to the present invention, these compounds can be produced efficiently.

  The whitening agent and antioxidant used in the present invention can be obtained by organic synthesis or extraction and purification from specific fungi, but are not limited to these methods.

  The active ingredients of the whitening agent and the antioxidant of the present invention can be obtained from the mycelium culture medium of Horotake mushroom by the following method, but are not limited thereto.

  The mycelium of Hourotake mushroom is a cell body cultured aseptically on a culture medium from a wild or artificially grown fruit body or mycelium itself, or a cell body preserved by means of subculture, freezing, freeze-drying, drying, etc. Can be used.

  Any fungus body may be used. For example, it is possible to use cells stored in storage facilities in various parts of the world, as well as cells that are native not only in Japan but also in various parts of the world. In addition, it can be used in the same manner as long as it is a closely related bacterial species. Artificially cultivated fruit bodies can also be used.

  The medium used for culturing the mycelium is potato dextrose agar medium, potato sucrose agar medium, peptone dextrose agar medium, and other agar medium, potato dextrose medium, potato sucrose medium, peptone dextrose medium, and other liquid medium, Any medium can be used as long as it can be used for culturing fungi. The mycelium culture method, culture conditions, etc. are not particularly limited, but a mycelium sufficiently grown by culturing by stationary culture, aeration culture, shaking culture, etc. at 20-30 ° C. for 10-50 days should be used. desirable.

  When the mycelium is induced from a wild fruit body or an artificially grown fruit body, it is induced to the mycelium by culturing aseptically from the tissue inside the fruit body or the spores of the fruit body on the above medium. Can do. Moreover, if it is a microbial cell already isolated and preserve | saved by means, such as a subculture, freezing, freeze-drying, and drying, it can culture on said medium as it is.

  The compound used in the present invention can be obtained by separating or purifying the mycelium from the fungus mycelia or the mycelium itself by extraction or various chromatography. The method for purifying or isolating the compound of the present invention from the mycelium of Butter mushroom is not particularly limited. In any method, the following structural formula can be obtained by hydrogenation from daedalin A obtained by extracting and purifying from mycelia of Horotake mushroom or from a fraction containing the compound shown in daedalin A at a high concentration.

に表される化合物Aを誘導し、美白剤および抗酸化剤として使用することができる。

And can be used as a whitening agent and an antioxidant.

  The method for adding hydrogen to daedalinA is not particularly limited. As a preferred production method, hydrogen is added to daedalin A in a yield of almost 100% by dissolving daedalin A in an organic solvent or water, mixing palladium carbon and hydrogen gas, and reacting at room temperature for several hours. Compound A can be obtained.

  In addition, the whitening agent and the antioxidant of the present invention may be a racemic mixture of daedalin A obtained by simple means such as organic synthesis or a racemic mixture of compound A obtained by adding hydrogen to daedalin A. Can be used as

  The method for producing the racemic mixture of daedalin A and the racemic mixture of compound A used in the present invention is not particularly limited.

When a racemic mixture of daedalin A is synthesized by the production method of the present invention, in a preferred embodiment of the present invention, the following steps are performed:
(1) The following structural formula used for manufacturing

を有する化合物１を提供する工程、
（２）化合物1とメタクロロパーベンゾイックアシッド（ｍＣＰＢＡ）とを反応させ、以下の構造式：

Providing compound 1 having:
(2) Compound 1 is reacted with metachloroperbenzoic acid (mCPBA) to give the following structural formula:

を有する化合物２を調製する工程、
（３）上記の化合物２から、以下の構造式：

Preparing compound 2 having
(3) From the above compound 2, the following structural formula:

で表される化合物３を調製する工程、
を包含する調製法が行われる。

A step of preparing compound 3 represented by:
A preparation process is carried out.

The manufacturing method of the compound 1 used for the manufacturing method of this invention is not specifically limited. As a preferable production method, 2,5-dihydroxybenzaldehyde is dissolved in a 50% THF / H ₂ O solution, and indium and 3-chloro-2-methyl-1-propene are added to obtain a reaction mixture. Although reaction time is not specifically limited, Preferred reaction time is 1-36 hours, More preferably, it is 6-24 hours. Thereafter, Compound 1 can be separated from the reaction mixture by separation techniques such as filtration, extraction, solvent fractionation, and various chromatography. Further, compound 1 can be used for the next synthesis reaction without separation.

  2,5-dihydroxybenzaldehyde used in the production method of the present invention is registered as Cas No. 1194-98-5. As 2,5-dihydroxybenzaldehyde, those commercially available from reagent manufacturers can be used as they are.

  Compound 2 can be synthesized by dissolving Compound 1 thus obtained in dichloromethane and reacting with metachloroperbenzoic acid. Although reaction temperature is not specifically limited, The preferable reaction temperature is 0-5 degreeC. The reaction time is not particularly limited, but a preferable reaction time is 1 to 4 hours. Thereafter, the compound 2 can be separated from the reaction mixture by separation techniques such as filtration, extraction, solvent fractionation, and various chromatography. In addition, compound 2 can be used in the next synthesis reaction without separation.

  Dichloromethane used in the production method of the present invention is registered as Cas number 75-09-2. As dichloromethane, those commercially available from reagent manufacturers can be used as they are.

  The metachloroperbenzoic acid used in the production method of the present invention is registered as Cas number 937-14-4. As the metachloroperbenzoic acid, those commercially available from reagent manufacturers can be used as they are.

  Compound 3 ((±) -daedalinA) can be obtained in a racemic form by dissolving compound 2 thus obtained in 1,2-dichloroethane and adding a catalytic amount of paratoluenesulfonic acid to react. it can. Although reaction temperature is not specifically limited, A preferable reaction temperature is 60-100 degreeC. The reaction time is not particularly limited, but a preferable reaction time is 2 to 8 hours. Thereafter, the compound 3 can be separated from the reaction mixture by separation techniques such as filtration, extraction, solvent fractionation, and various chromatography.

  The whitening agent and antioxidant of the present invention can be used as an external preparation for skin, foods and pharmaceuticals. The shape of the external preparation for skin, food and medicine is not particularly limited. Any shape can be selected depending on the administration method. For example, it can be used in the form of liquid, gel, cream, granule, solid, gas such as aerosol. When used as a whitening agent and an antioxidant of the present invention, lotion, cream, gel, ointment, milky lotion, serum, pack, face wash, cleansing agent, hair care agent, soap, bath preparation, shampoo, rinse, lipstick , Lipstick, foundation, and other cosmetics, quasi drugs, and pharmaceuticals.

  Skin external preparations containing the whitening agent and antioxidant of the present invention are oils, waxes, hydrocarbons, silicones, fatty acids, alcohols, esters, surfactants, thickeners, powders, etc. In addition to cosmetic base materials, active ingredients of pharmaceuticals and quasi-drugs, pH adjusters, preservatives, pigments, fragrances, antioxidants, natural extracts, and the like can be blended as necessary.

  Well-known means can be used as a method for preparing the compound of the present invention as a skin external preparation. Representative preparation methods are illustrated below, but are not limited thereto.

(Preparation of cosmetic cream formulation)
Cosmetic creams can be formulated using the following ingredients and recipe:
(A) stearic acid 8.0 (wt%),
(B) stearyl alcohol 4.0 (wt%),
(C) butyl stearate 6.0 (wt%),
(D) glyceryl monostearate 2.0 (% by weight),
(E) propylene glycol 5.0 (% by weight),
(F) Potassium hydroxide 0.4 (wt%),
-(G) antiseptic (paraoxybenzoic acid ester) 0.1-0.5 (wt%),
-(H) perfume 0.001-0.1 (wt%),
(I) antioxidant (dibutylhydroxytoluene) 0.05 to 0.15 (% by weight),
(J) ethanol 5.0 (% by weight),
(K) an effective amount of a compound of the invention,
(L) Purified water added as the balance above.
(Manufacturing method)
1. A part of the above (A) to (D), (E) to (I), and (L) is separately heated and mixed, mixed at 80 ° C., stirred with a homomixer, and then cooled to 50 ° C. .
2. (J) is added to a part of (L), and (K) is dissolved.
3. The above solution 1 and solution 2 are mixed and cooled to 30 ° C.

(Preparation of emulsion formulation)
An emulsion can be formulated using the following ingredients and recipe:
(A) stearic acid 2.0 (wt%),
(B) Cetyl alcohol 1.5 (% by weight),
-(C) Vaseline 4.0 (wt%),
(D) Squalane 5.0 (% by weight),
(E) Glycerol tri-2-ethylhexanoate 2.0 (wt%),
(F) sorbitan monooleate 2.0 (wt%),
(G) Glycerol 9.0 (% by weight),
(H) potassium hydroxide 0.1 (% by weight),
(I) Preservative (paraoxybenzoic acid ester) 0.1 to 0.5 (% by weight),
・ (J) Fragrance 0.001-0.1 (% by weight)
(K) antioxidant (dibutylhydroxytoluene) 0.05 to 0.15 (% by weight),
-(L) Ethanol 5.0 (wt%)
(M) an effective amount of a compound of the invention,
(N) Purified water added as the balance above.
(Manufacturing method)
1. A part of (A) to (F), (G) to (K), and (N) is separately heated and mixed, mixed at 80 ° C, stirred with a homomixer, and then cooled to 50 ° C.
2. (L) is added to a part of (N), and (M) is dissolved.
3. The above solution 1 and solution 2 are mixed and cooled to 30 ° C.

(Preparation of lotion)
Lotion can be formulated using the following ingredients and recipe:
(A) POE (20) oleyl alcohol ether 0.5 (% by weight)
(B) glycerin 10 (% by weight),
(C) methylcellulose 0.2 (% by weight),
-(D) Preservative (paraoxybenzoic acid ester) 0.1-0.5 (wt%),
-(E) perfume 0.001-0.1 (wt%),
(F) Antioxidant (dibutylhydroxytoluene) 0.05 to 0.15 (% by weight),
・ (G) Ethanol 5.0 (% by weight)
(H) an effective amount of a compound of the invention,
(I) Purified water added as the balance above.
(Manufacturing method)
The above (A) to (I) are uniformly dissolved.

(Preparation of pack agent)
The pack can be formulated using the following ingredients and recipe:
(A) Glycerin 12 (% by weight),
(B) Montmorillonite 10.0 (wt%),
(C) Titanium oxide 8.0 (wt%),
(D) Kaolin 10.0 (wt%),
(E) Preservative (paraoxybenzoic acid ester) 0.1 to 0.5 (% by weight),
-(F) Fragrance 0.001-0.1 (wt%),
(G) antioxidant (dibutylhydroxytoluene) 0.05 to 0.15 (% by weight),
・ (H) Ethanol 5.0 (% by weight)
(I) an effective amount of a compound of the invention,
-(J) Purified water added as the balance above.
(Manufacturing method)
The above (A) to (J) are uniformly dissolved.

  When the whitening agents and antioxidants of the present invention are used as pharmaceuticals, they can be formulated in combination with a pharmaceutically acceptable carrier type (eg, sterile carrier). “Pharmaceutically acceptable carrier” refers to a non-toxic individual or liquid filler, diluent, encapsulant, or any type of formulation adjuvant.

  When a composition containing a compound of the present invention is used as a medicine, it takes into account individual individuals, administration method, administration schedule and factors known to those skilled in the art, and adheres to medical practice standards (GMP = good medical practice) Formulate and dose in a manner that Accordingly, in the present specification, the “effective amount” is determined by taking such consideration into consideration.

  Therapeutic agents can be administered orally, rectally, parenterally, in the bath, vaginally, intrathecally, topically (such as by powders, ointments, gels, drops, or transdermal patches), orally or orally or nasally Can be administered as a spray. As used herein, the term “parenteral” refers to modes of administration including intravenous and intramuscular, intraperitoneal, intrasternal, subcutaneous and indirect injections and infusions.

  The present invention will be described below based on examples, but the following examples are provided for illustrative purposes only. Accordingly, the scope of the present invention is not limited to the above detailed description of the invention nor the following examples, but is limited only by the scope of the claims.

(Example 1: Cultivation of mycelium from fruit body of Horotake mushroom)
The fruit body of Horotakutake, which grows naturally in Japan, was collected, and the internal tissue was aseptically cut out from the fruit body. This tissue was transferred to a peptone dextrose agar plate and cultured at 25 ° C. for 20 days. Twenty days later, the grown mycelium of Butter mushrooms was transferred to a new agar medium. This mycelium obtained by repeating this operation twice was cultured in a peptone dextrose liquid medium (10 l) for 40 days at 25 ° C. After 40 days, the mycelium was filtered off from the culture broth after culturing, and the mycelium was removed. The cultivated mycelium of Horotakutake mycelium was concentrated under reduced pressure to obtain a concentrate (50 g). The composition of peptone dextrose agar and peptone dextrose liquid medium is shown below.

(Composition of peptone dextrose agar)
Polypeptone: 5g
Glucose: 20g
Yeast extract: 2g
Magnesium sulfate heptahydrate: 0.5g
Potassium dihydrogen phosphate: 1g
Powder agar: 20g
Water: 1000g.

(Composition of peptone dextrose liquid medium)
Polypeptone: 5g
Glucose: 30g
Yeast extract: 2g
Magnesium sulfate heptahydrate: 0.5g
Potassium dihydrogen phosphate: 1g
Water: 1000g.

(Example 2: Purification of active substance from culture filtrate of Horotake mushroom mycelium)
An ethyl acetate elution fraction (5 g) was obtained by partitioning 50 g of the culture filtrate concentrate of Horotake mushroom mycelium with ethyl acetate and water. The fraction eluted with ethyl acetate was separated by silica gel chromatography using Wakogel C300, 150 g as silica gel. As the elution solvent, 450 ml each of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% ethyl acetate / hexane elution solvent was used. Separated into minutes.

  Among the obtained fractions, the 50% ethyl acetate / hexane fraction (0.85 g) was further separated by silica gel chromatography using Wakogel C300, 160 g as silica gel. Separation and purification using 30% acetone / hexane as the elution solvent

に示す化合物ｄａｅｄａｌｉｎＡを３８４ｍｇの収率で得た。

The compound daedalin A shown in the above was obtained in a yield of 384 mg.

(Example 3: Measurement of spectral data)
For reference, mass spectrometry (MS), optical rotation, infrared absorption spectrum (IR), ultraviolet absorption spectrum (UV), and nuclear magnetic resonance spectrum ( ¹ H-NMR, ¹³ C-NMR) spectrum data of daedalin A are shown below. .

(Spectral data)
Property: yellow oily substance mass spectrometry: 191.0712 [M ⁺ -H] (calculated value 191.0708: C ₁₁ H ₁₁ O ₃ )
Optical rotation: [α] ²⁹ _D + 15.4 ° (c1.49, EtOH)
Ultraviolet absorption spectrum: 263 nm, ε: 2.6 × 10 ³ , λ: 333 nm, ε: 2.4 × 10 ³
Infrared absorption spectrum (cm ⁻¹ ): 3389, 2928, 2360, 2342, 1490, 1459, 1227, 1051, 816, 764, 711
Nuclear magnetic resonance spectrum ( ¹ H-NMR, 500.1 MHz, CDCl ₃ ): δ 6.67 (1H, d, J = 8.6 Hz), δ 6.60 (1 H, dd, J = 2.9 Hz, 8.6 Hz) ), Δ 6.50 (1H, d, J = 2.9 Hz), δ 6.38 (1H, d, J = 9.9 Hz), δ 5.61 (1H, d, J = 9.9 Hz), δ 3.68. (1H, d, J = 11.6 Hz), δ3.59 (1H, d, J = 11.6 Hz), δ1.35 (1H, s)
Nuclear magnetic resonance spectrum ( ¹³ C-NMR, 125.8 MHz, CDCl 3): δ 149.8 (C), δ 146.8 (C), δ 128.0 (CH), δ 124.7 (CH), δ 121.7 (C ), Δ 116.8 (CH), δ 115.8 (CH), δ 113.2 (CH), δ 79.0 (C), δ 68.5 (CH ₂ ), δ 22.3 (CH ₃ )
These spectral data agreed with the spectral data of daedalin A isolated so far.

(Example 4: Hydrogenation to dadalin A)
21 mg (0.11 mmol) daedalin A was dissolved in ethanol and mixed with palladium carbon (0.1 mg, 0.01 mmol) and hydrogen gas. After 8 hours, the palladium carbon was removed and the resulting ethanol solution was concentrated. This concentrate was subjected to silica gel chromatography using 50% ethyl acetate / hexane as an elution solvent to obtain the following chemical formula.

に示す化合物Ａを１００％の収率で得た。

Was obtained in a yield of 100%.

(Example 5: Measurement of spectral data)
For reference, the spectrum data of Compound A is shown below.
Property: colorless oily mass analysis: HREIMS: 194.0949 (theoretical value C ₁₁ H ₁₄ O ₃ , 194.0943))
Optical rotation: [α] ¹⁹ _D −9.9 ° (c0.21, Acetone)
Infrared absorption spectrum (cm ⁻¹ ): 3353, 2930, 1618, 1494, 1454, 1351, 1285, 1222, 1152, 1097, 1050, 923, 898, 809, 737, 705
Nuclear magnetic resonance spectrum ( ¹ H-NMR, 500.1 MHz, CDCl ₃ ): δ6.66 (1H, d, J = 8.7 Hz), δ6.59 (1H, dd, J = 2.9 Hz, 8.7 Hz) ), Δ6.56 (1H, d, J = 2.9 Hz), δ5.40 (1H, brs), δ3.64 (1H, d, J = 12 Hz), δ3.60 (1H, d, J = 12 Hz) ), Δ 2.79 (1H, ddd, J = 6.3 Hz, 11 Hz, 14 Hz), δ 2.68 (1 H, dt, J = 5.2 Hz, 17 Hz), δ 2.27 (1 H, brs), δ 1.9. (1H, ddd, J = 6.1 Hz, 11 Hz, 14 Hz), δ 1.67 (1H, ddd, J = 4.5 Hz, 6.2 Hz, 14 Hz), δ 1.24 (3H, s)
Nuclear magnetic resonance spectrum ( ¹³ C-NMR, 125.8 MHz, CDCl 3): δ 149.0, δ 147.1, δ 121.9, δ 117.7, δ 115.4, δ 114.6, δ 76.2, δ 69.1, δ 27 .5, δ21.8, δ20.4.

(Example 6: Confirmation of stability of Compound A)
When Compound A was adjusted to 1 mg / ml and stored in 50% ethanol aqueous solution for 6 months at 25 ° C., the compound A solution showed no change such as coloring. On the other hand, when daedalin A was stored under the same conditions for the same period, yellow color was observed in daedalin A. Therefore, Compound A exhibits better stability than daedalin A in solution.

(Example 7: Tyrosinase inhibition test using tyrosine as a substrate)
In order to examine the whitening effect of Compound A, a tyrosinase inhibition test was performed on Compound A. In this test, tyrosine was used as a reaction substrate. The tyrosinase inhibition test was carried out by the following test method. The tyrosinase inhibitory activity of Compound A using tyrosine as a substrate was compared with arbutin and daedalin A which are commercially available as whitening agents.

(Test method)
1. In phosphate buffer (PH 6.8), tyrosinase was dissolved in 40 U / ml and L-tyrosine as 1 mM as final addition amounts, and the test substance was dissolved in dimethyl sulfoxide (DMSO) and mixed.
2. After mixing, it was incubated at 37 ° C. for 60 minutes.
The tyrosinase inhibition rate was calculated by substituting the absorbance value at 3.475 nm into the following equation.
Tyrosinase inhibition rate = 100 × {(A−B) − (C−D)} / (A−B)
In this formula, A, B, C, and D mean the following.
A: Absorbance 60 minutes after reaction of control (no test substance added) B: Absorbance value before reaction of control (no test substance added) C: Absorbance value 60 minutes after reaction of test substance added sample D: Test Absorbance value before reaction of sample with added substance.

  Table 1 below shows the tyrosinase inhibitory activity for each added amount of Compound A, daedalin A, and arbutin.

いずれの化合物も０．４ｍＭ，０．２ｍＭ，０．１ｍＭ，０．０５ｍＭの添加量で濃度依存的にチロシナーゼの活性を阻害した。５０％阻害活性を示す添加濃度であるＩＣ_５０を計算すると、それぞれ化合物Ａは２８９μＭ，ｄａｅｄａｌｉｎＡは１９４μＭ，アルブチンは６７２μＭであった。ＩＣ_５０で比較すると化合物Ａのチロシナーゼ阻害作用はアルブチンよりも強いが、ｄａｅｄａｌｉｎＡよりも少し弱い阻害作用であった。本試験においてｄａｅｄａｌｉｎＡはチロシナーゼの添加により黄色に変色するが、化合物Ａには変色は認められず、チロシナーゼに対する安定性はｄａｅｄａｌｉｎＡよりも化合物Ａの方が優れていることを確認した。

All compounds inhibited the activity of tyrosinase in a concentration-dependent manner at the addition amounts of 0.4 mM, 0.2 mM, 0.1 mM and 0.05 mM. When IC ₅₀ , which is an addition concentration showing 50% inhibitory activity, was calculated, Compound A was 289 μM, daedalin A was 194 μM, and arbutin was 672 μM. When compared with IC ₅₀ , the tyrosinase inhibitory effect of Compound A was stronger than that of arbutin, but slightly weaker than daedalinA. In this test, daedalin A turns yellow with the addition of tyrosinase, but no color change was observed in compound A, and it was confirmed that compound A was superior to daedalin A in terms of stability to tyrosinase.

(Example 8: Tyrosinase inhibition test using DOPA as a substrate)
A tyrosinase inhibition test was then performed using DOPA as the reaction substrate. By performing a tyrosinase inhibition test using DOPA, it is possible to measure the inhibitory action of a presentation substance on the oxidation reaction from DOPA to DOPAquinone in melanin synthesis. The tyrosinase inhibitory action of Compound A in this test was compared with daedalin A and Compound A.

(Test method)
1. In phosphate buffer (PH 6.8), tyrosinase was dissolved to a final addition amount of 20 U / ml and L-DOPA was 1 mM, and the test substance was dissolved in DMSO and mixed.
2. After mixing, it was incubated at 25 ° C. for 30 minutes.
The tyrosinase inhibition rate was calculated by substituting the absorbance value at 3.475 nm into the following equation.
Tyrosinase inhibition rate = 100 × {(A−B) − (C−D)} / (A−B)
In this formula, A, B, C, and D mean the following.
A: Absorbance 60 minutes after the reaction of the control (no test substance added) B: Absorbance value before the reaction of the control (no test substance added) C: Absorbance value 30 minutes after the reaction of the test substance added sample D: Test Absorbance value before reaction of sample with added substance.

  Table 2 below shows tyrosinase inhibitory activity for each addition amount of Compound A, daedalin A, and arbutin.

本試験においては化合物Ａだけが０．２ｍＭ，０．４ｍＭ，０．８ｍＭの添加濃度で濃度依存的にチロシナーゼ活性を阻害したが、ｄａｅｄａｌｉｎＡとアルブチンは全くチロシナーゼ阻害作用を示さなかった。従って、化合物ＡだけがＤＯＰＡが基質であっても、これらの添加濃度においてチロシナーゼ活性を阻害した。従って、化合物ＡはｄａｅｄａｌｉｎＡやアルブチンよりも効果的な美白作用を示すことが期待できる。

In this test, only Compound A inhibited tyrosinase activity in a concentration-dependent manner at addition concentrations of 0.2 mM, 0.4 mM, and 0.8 mM, but daedalin A and arbutin did not show any tyrosinase inhibitory action. Thus, only Compound A inhibited tyrosinase activity at these added concentrations even though DOPA was a substrate. Therefore, compound A can be expected to show a whitening effect more effective than daedalin A or arbutin.

(Example 9: DPPH radical scavenging test of compound A)
In order to investigate the antioxidant effect of Compound A, DPPH radical scavenging test was performed on Compound A. The DPPH radical scavenging action of Compound A was compared with daeadalin A and Trolox marketed as a strong antioxidant.

(Test method)
1. DPPH was dissolved in ethanol and adjusted to 0.1 mmol.
2. The test substance dissolved in 50% ethanol aqueous solution was dissolved in Tris-HCl buffer (PH7.4).
3. The above 1 and 2 solutions were mixed 1: 1 and allowed to react for 20 minutes at 25 ° C. in the dark.
The DPPH radical scavenging rate was calculated by substituting the absorbance value of 4.517 nm into the following equation.
DPPH radical elimination rate = 100 × (A−B) / (C−D)
In this formula, A, B, C, and D mean the following.
A: Absorbance value after reaction of test substance-added sample B: Absorbance value when ethanol is added instead of DPPH of the test substance-added sample.
C: Absorbance value after reaction of control (no test substance added).
D: Absorbance value when ethanol was added instead of DPPH as a control (no test substance added).

  Table 3 shows the DPPH radical scavenging rate for each additive concentration of Compound A, daedalin A, and Trolox.

化合物A のDPPHラジカル消去作用はdaedalin A とほぼ同等であり，トロロックス よりも若干弱いが十分なラジカル消去作用であった。化合物A は十分な効果と安定性を持つ優れた抗酸化剤である。

The DPPH radical scavenging action of Compound A was almost the same as that of daedalin A, which was a little weaker than Trolox but was a sufficient radical scavenging action. Compound A is an excellent antioxidant with sufficient efficacy and stability.

(Example 10: Racemic form of daedalin A (chemical synthesis of (±) -daedalin A))
(±) -daedalinA can be synthesized by the following synthesis method.
To a solution of 2,5-dihydroxybenzaldehyde (22.5 mg, 0.16 mmol) in THF / H ₂ O (1/1, 6.0 ml) was added indium (18.4 mg, 0.16 mmol) and 3-chloro-2- Methyl-1-propene (0.05 ml, 0.48 mmol) was added sequentially. After stirring at room temperature for 12 hours, the reaction was stopped by adding saturated brine, and the mixture was extracted with ethyl acetate. The ethyl acetate extract was dried over magnesium sulfate, filtered, and concentrated under reduced pressure to remove the solvent. The concentrate was purified by silica gel column chromatography using a 50% ethyl acetate / hexane solution to obtain Compound 1 (31.6 mg, 0.16 mmol). A scheme for synthesizing Compound 1 is shown below.

次に，化合物１（３１．６ｍｇ，０．１６ｍＭ）のジクロロメタン溶液にｍＣＰＢＡ（８５．０ｍｇ，０．３２ｍｍｏｌ）を加え、室温にて２時間撹拌した。半飽和Ｎａ_２Ｓ_２Ｏ_３水溶液を加えて反応を停止し、ジエチルエーテルを用いて抽出した。抽出したジエチルエーテル溶液を飽和ＮａＨＣＯ_３水溶液、飽和食塩水で順次洗浄後、硫酸マグネシウムで乾燥し、ろ過した後、減圧濃縮することで溶媒を除去し，化合物２を含む混合物を得た。この濃縮物を精製することなく次の反応に使用した。下記に化合物２を合成するためのスキームを示す。

Next, mCPBA (85.0 mg, 0.32 mmol) was added to a dichloromethane solution of compound 1 (31.6 mg, 0.16 mM), and the mixture was stirred at room temperature for 2 hours. Half-saturated Na ₂ S ₂ O ₃ aqueous solution was added to stop the reaction, and the mixture was extracted with diethyl ether. The extracted diethyl ether solution was washed successively with saturated aqueous NaHCO ₃ solution and saturated brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to remove the solvent and obtain a mixture containing compound 2. This concentrate was used in the next reaction without purification. A scheme for synthesizing Compound 2 is shown below.

１，２−ジクロロエタンに化合物２を含む濃縮物を溶解し、触媒量のパラトルエンスルホン酸を加え、８３℃にて６時間撹拌した。反応液を室温まで冷却後、飽和ＮａＨＣＯ_３水溶液にて反応を停止し、酢酸エチルにて抽出した。酢酸エチル抽出液を飽和食塩水で洗浄後，硫酸マグネシウムで乾燥し，ろ過した後、減圧濃縮して溶媒を除去した。この濃縮物を５０％酢酸エチル／ヘキサン溶液によるシリカゲルカラムクロマトグラフィーにより精製し、（±）−ｄａｅｄａｌｉｎＡ（１１．０ｍｇ，０．０５７ｍｍｏｌ）を得た。化合物１から２段階の反応における（±）−ｄａｅｄａｌｉｎＡの反応収率は３５％であった。（±）−ｄａｅｄａｌｉｎＡを合成するためのスキームは、

A concentrate containing Compound 2 was dissolved in 1,2-dichloroethane, a catalytic amount of para-toluenesulfonic acid was added, and the mixture was stirred at 83 ° C. for 6 hours. The reaction mixture was cooled to room temperature, quenched with saturated aqueous NaHCO ₃ solution, and extracted with ethyl acetate. The ethyl acetate extract was washed with saturated brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to remove the solvent. The concentrate was purified by silica gel column chromatography using a 50% ethyl acetate / hexane solution to obtain (±) -daedalin A (11.0 mg, 0.057 mmol). The reaction yield of (±) -daedalinA in the two-step reaction from compound 1 was 35%. The scheme for synthesizing (±) -daedalinA is:

である。

It is.

(Example 11: Measurement of spectral data of compound 1)
For reference, the spectrum data of Compound 1 is shown below.
Property: Brown crystal mass spectrometry: HREIMS: 194.0925 (theoretical value C ₁₁ H ₁₄ O ₃ , 194.0943)
Infrared absorption spectrum (cm ⁻¹ ): 3388, 2929, 1454, 1419, 1257, 1086, 1047
Nuclear magnetic resonance spectrum ( ¹ H-NMR, 500.1 MHz, CDCl ₃ ): δ1.31 (1H, s), δ1.83 (3H, s), δ2.45 (1H, dd, J = 3.5, 14 Hz), δ 2.61 (1H, dd, J = 10.5, 14 Hz), δ 4.47 (1 H, d, J = 3.5 Hz), δ 4.87 (1 H, dd, J = 2.5, 10 .5 Hz), δ4.92 (1H, s), δ5.01 (1H, s), δ6.52 (1H, d, J = 3.0 Hz), δ6.66 (1H, dd, J = 3.0) , 8.5 Hz), δ 6.75 (1H, d, J = 8.5 Hz), δ 7.61 (1H, s)
Nuclear magnetic resonance spectrum ( ¹³ C-NMR, 125.8 MHz, CDCl ₃ ): δ 149.4, δ 148.5, δ 141.7, δ 127.2, δ 118.0, δ 115.5, δ 115.1, δ 113.7, δ 72.4, δ 46.2, δ 22.2.

(Example 12: Measurement of spectral data of (±) -daedalinA)
For reference, the spectrum data of Compound 3 ((±) -daedalinA) is shown below.
Properties: colorless oily mass analysis: HREIMS: 192.0754 (theoretical value C ₁₁ H ₁₄ O ₃ , 192.0786)
Infrared absorption spectrum (cm ⁻¹ ): 3337, 2979, 1489, 1459, 1223, 1047
Nuclear magnetic resonance spectrum ( ¹ H-NMR, 500.1 MHz, CDCl ₃ ): δ 1.36 (3H, s), δ 3.60 (1 H, d, J = 11.5 Hz), δ 3.68 (1 H, d, J = 11.5 Hz), δ 5.62 (1 H, d, J = 10.0 Hz), δ 6.39 (1 H, d, J = 10.0 Hz), δ 6.50 (1 H, d, J = 3.0 Hz) ), Δ 6.60 (1H, dd, J = 3.0 Hz), δ 6.67 (1H, d, J = 8.5 Hz)
Nuclear magnetic resonance spectrum ( ¹³ C-NMR, 125.8 MHz, CDCl ₃ ): δ 149.8, δ 146.1, δ 127.9, δ 124.7, δ 121.7, δ 116.8, δ 115.8, δ 113.2, δ 79.0, δ 68.5, δ 22.3.

(Example 13 Yield of chemical synthesis of (±) -daedalinA by reaction solvent)
In order to examine the synthesis yield of (±) -daedalinA by the reaction solvent, (±) -daedalinA was synthesized from compound 1 using benzene, toluene, dichloromethane, or dichloroethane as the reaction solvent. The synthesis was performed by the method described in Example 10. The reaction yield when (±) -daedalin A was synthesized from compound 1 for each reaction solvent was compared. The results are shown in the table below.

ベンゼンを使用した場合の反応収率は０．５％、トルエンを使用した場合の反応収率は０．６％、ジクロロメタンを使用した場合の反応収率は０．５％と、ジクロロエタンを使用した場合の反応収率（３５％）よりも大きく劣り、実用にはそぐわない。

The reaction yield when using benzene was 0.5%, the reaction yield when using toluene was 0.6%, the reaction yield when using dichloromethane was 0.5%, and dichloroethane was used. It is much inferior to the reaction yield (35%) in this case and is not suitable for practical use.

  Therefore, (±) -daedalinA can be obtained in a high yield by using 1,2-dichloroethane as a reaction solvent in the reaction for obtaining dadealinA from compound 2.

(Example 14: (±) -daedalinA tyrosinase inhibition test)
In order to examine the whitening effect of (±) -daedalinA, a tyrosinase inhibition test was performed on (±) -daedalinA. In this test, tyrosine was used as a reaction substrate. The tyrosinase inhibition test was carried out by the test method shown in Example 7. The tyrosinase inhibitory action of (±) -daedalin A was compared with arbutin and daedalin A which are commercially available as whitening agents.

  Table 4 below shows tyrosinase inhibitory activity for each addition amount of Compound A, daedalin A, and arbutin.

本試験から、(±)-daedalinA は市販の美白剤であるアルブチン以上のチロシナーゼ阻害作用を示すと共に、そのチロシナーゼ阻害活性はdaedalin A とほぼ同等であった。従って、簡便な合成法による製造法によって製造できるラセミ体の(±)-daedalin A であっても、光学活性を持つdaedalin A の代替使用が可能である。

From this test, (±) -daedalinA showed a tyrosinase inhibitory activity higher than that of arbutin, which is a commercially available whitening agent, and its tyrosinase inhibitory activity was almost equivalent to daedalin A. Therefore, even racemic (±) -daedalin A that can be produced by a production method based on a simple synthesis method can be used instead of daedalin A having optical activity.

(Example 15: DPPH radical elimination rate measurement test of (±) -daedalin A)
In order to investigate the antioxidant action of (±) -daedalinA, a DPPH radical scavenging rate measurement test was performed on (±) -daedalinA. The DPPH radical scavenging action of Compound A was compared with daeadalin A and Trolox marketed as a strong antioxidant.

本試験から、（±）−ｄａｅｄａｌｉｎＡの抗酸化作用はｄａｅｄａｌｉｎＡとほぼ同等であった。従って、簡便な合成法による製造法によって製造できるラセミ体の（±）−ｄａｅｄａｌｉｎＡであっても、光学活性を持つｄａｅｄａｌｉｎＡの代替使用が可能である。

From this test, the antioxidant action of (±) -daedalinA was almost the same as daedalinA. Therefore, even racemic (±) -daedalin A that can be produced by a production method based on a simple synthesis method can be used instead of daedalin A having optical activity.

(Example 15: Measurement of cultivation period of boletus mushroom and production amount of daedalin A)
The relationship between the cultivation time of edible mushrooms and the production amount of daedalin A was investigated by HPLC analysis.
(Method)
(I) The mycelium of Horotake mushrooms separated by the method described in Example 1 was statically cultured at 25 ° C. in the peptone dextrose liquid medium shown in Example 1.
(Ii) Every 10 days after the start of cultivating Horotake mushroom, the culture filtrate of Horotake mushroom was sampled.
(Iii) The culture filtrate was diluted to 10% with a 50% aqueous methanol solution and filtered through a membrane filter.
(Iv) The culture filtrate prepared as described above was subjected to HPLC analysis under the following analysis conditions.
(V) The content (mg) of daedalinA contained in 1 L of culture broth by the mycelium of Borotake mushroom was calculated by comparing the retention time and absorbance of HPLC analysis of daedalinA (1 mg / ml).
(Vi) The production amount of daedalin A in this test was calculated by adding coumarin (1 mg / ml) as an internal standard substance and using this peak as a standard.
(Analysis conditions)
1) Column: ODS column, inner diameter 4.6 mm, length 150 mm, particle size 5 μm
2) Mobile phase: 50% methanol / water 3) Flow rate: 0.5 ml / min
4) Detector: UV333nm
5) Temperature: 30 ° C
6) Loop: 20 μl
7) Sample: 5 ml charged.

(result)
The results of the above test are shown in the following table.

ホウロクタケ菌糸体による daedalin A の生産量を培養期間毎に測定すると、ホウロクタケは 30 日まで培養期間が経過すると daedalin A の生産量は増加するが、30日目以降は時間の経過とともにdaedalin A の含有量は減少する。

When the amount of daedalin A produced by the mycelium of borochoraceae is measured at each culture period, the amount of daedalin A increases as the culture period elapses until the 30th day, but the content of daedalinA increases with the passage of time from the 30th day. The amount decreases.

  From the above results, it is considered that daedalinA is a substance that is produced along with the growth of mycelia of Horotake mushrooms and then metabolized by mycelial growth.

  Therefore, in order to efficiently produce daedalin A, it is necessary to rapidly isolate daedalin A from the mycelium after culturing the mycelium of Borotake mushroom for a certain period and before it is degraded in the metabolic process of Borotake mycelium .

  As mentioned above, although this invention has been illustrated using preferable embodiment of this invention, this invention should not be limited and limited to this embodiment. It is understood that the scope of the present invention should be construed only by the claims. It is understood that those skilled in the art can implement an equivalent range based on the description of the present invention and the common general technical knowledge from the description of specific preferred embodiments of the present invention. Patents, patent applications, and documents cited herein should be incorporated by reference in their entirety, as if the contents themselves were specifically described herein. Understood.

  By adding hydrogen to daedalin A contained in the cultured metabolite of the mycelium of Horotake mushroom, it is possible to provide an antioxidant and a whitening agent that are more stable.

  In addition, it has been clarified that even racemic (±) -daedalin A prepared by the synthesis method of the present invention exhibits sufficient antioxidant and whitening effects, and efficient (±) -daedalinA and daedalin A Clarified the manufacturing method of A.

Claims (10)

The following structural formula:

The whitening agent containing the compound which has this. The following structural formula:

An antioxidant containing a compound having The following structural formula:

An active oxygen scavenger containing a compound having: The following structural formula:

An external preparation for skin containing a compound having The structural formula of the following:

A whitening agent comprising a racemic mixture of the compound represented by The structural formula of the following:

An antioxidant comprising a racemic mixture of the compounds represented by The structural formula of the following:

An active oxygen scavenger containing a racemic mixture of the compounds represented by The structural formula of the following:

A skin external preparation containing a racemic mixture of the compound represented by The following structural formula:

A method for producing a compound represented by the following:
a) culturing mycelia of D aedalea dickinsii for 10 to 50 days;
b) From the culture of step (a), the following structural formula

Extracting a compound represented by: and
c) adding hydrogen to the compound extracted in step (b);
Including the method. The following structural formula:

A process for preparing a racemic mixture of the compounds represented by
(A) Paratoluenesulfonic acid and the following structural formula

A step of reacting the compound represented by: in 1,2-dichloroethane,
(B) adding hydrogen to the compound produced in step (a);
Including the method.